Detection of Human and Mouse CDK4 by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line, HeLa human cervical epithelial carcinoma cell line, and NIH‑3T3 mouse embryonic fibroblast cell line. PVDF Membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse CDK4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5254) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CDK4 at approximately 34 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
CDK4 in HeLa Human Cell Line. CDK4 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human/Mouse CDK4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5254) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
CDK4 (cyclin-dependent kinase 4) is a 34 kDa member of the CMGC Ser/Thr protein kinase family of enzymes. Within the cell, pRb/retinoblastoma protein is normally unphosphorylated and suppresses cell growth at the G1 stage of the cell cycle. When mitosis is required, CDK4 is phosphorylated, forms a complex with cyclin D1, 2 or 3, and phosphorylates pRb/retinoblastoma protein. This inactivates pRb and cells initiate DNA synthesis. Human CDK4 is 303 amino acids (aa) in length. It contains one protein kinase domain (aa 6‑295) and an activating phosphorylation site at Thr172. There are three potential variants; one that shows an alternate start site at Met75, a second that shows a two aa substitution for aa 89‑211, and a third that shows a 23 aa substitution for aa 89‑303. Full-length human and mouse CDK4 are 95% aa identical.
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